Talk:Amplex UltraRed: Difference between revisions
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In preliminary experiments Oroboros Instruments got indications that small amounts (as typically used in SUIT protocols) of the following substances are '''compatible''' with the method: | In preliminary experiments Oroboros Instruments got indications that small amounts (as typically used in SUIT protocols) of the following substances are '''compatible''' with the method: | ||
DMSO, ethanol, [[malate]], [[glutamate]], [[pyruvate]], [[succinate]], [[ADP]] + Mg2+, [[ATP]] + Mg2+, [[rotenone]], [[FCCP]], [[oligomycin]], [[antimycin A]], [[malonate]], [[myxothiazol]] | DMSO, ethanol, [[malate]], [[glutamate]], [[pyruvate]], [[succinate]], ([[ADP]] + Mg2+), ([[ATP]] + Mg2+), [[rotenone]], [[FCCP]], [[oligomycin]], [[antimycin A]], [[malonate]], [[myxothiazol]] | ||
Revision as of 09:39, 6 June 2012
Different Brands
| Trade Mark | Manufacturer/ Distributor | price [€/mg]
|
product id
|
| Amplex Red | 37.4
|
A12222
| |
| Amplex Ultra Red | 48.8
|
A36006
| |
| Ampliflu Red | 18.5
|
90101
| |
| Quanta Red | 15150
|
Conditions
| Citation | Amplex | HRP | pH | Limit of detection | |||
stock
|
work
|
unit definition
|
stock
|
work
|
|||
mM
|
µM
|
U/ml
|
U/ml
|
||||
| BIOTEK | 10
|
50
|
pyrogallol
|
10
|
0.1
|
7.4
|
4 nM (absorption 300 nm)
|
| INVITROGEN | 10
|
50
|
10
|
0.1
|
6 to 7.5
|
<80 nM
| |
| Towne 2004 | 160
|
0.41
|
7.5 to 8.5
|
100 nM
| |||
| Zhou1997 | 3 ?
|
0.3 to 1
|
50 nM (10 nm optimal cond)
| ||||
| Mohanty1997 | 100
|
10 to 100
|
1
|
18 nM
| |||
| Komary2010 | 1
|
2.5
|
|||||
At Oroboros Instruments we use the publication from Prof. Tretters OROBOROS MiPNet Reference Laboratory as a starting point for our development.
Substances incompatible with the Amplex Red method
The following substances/ classes of substances are strictly incompatible with the Amplex Red method for theoretical reasons:
- strongly redox active substances, e.g. cytochrome c, TMPD/Ascorbate
- inhibitors of horseradish peroxidase, see horseradish peroxidase
- catalase and other substances consuming H2O2. The effect of substances in the medium that consume H2O2 slowly is taken account of by the calibration procedure. However, such substances will decreases the sensitivity of the method.
The effect of other substances should be checked by blank experiments, including comparing the sensitivity (result of calibration) before and after injecting the substances. In preliminary experiments Oroboros Instruments got indications that small amounts (as typically used in SUIT protocols) of the following substances are compatible with the method:
DMSO, ethanol, malate, glutamate, pyruvate, succinate, (ADP + Mg2+), (ATP + Mg2+), rotenone, FCCP, oligomycin, antimycin A, malonate, myxothiazol
